【摘要】 通过农杆菌介导法将反义磷脂酶Dγ Anti-PLDγ 基因转入美洲黑杨G2中以提高其耐盐性.本研究建立了美洲黑杨G2组培再生体系,确立了美洲黑杨G2叶片分化最适宜培养基和生根培养基,进行了卡那霉素 选择性抗生素 敏感性试验,改良了传统的农杆菌介导转化法.经诱导不定芽及生根阶段卡那霉素 选择性抗生素 连续筛选,获得了21株卡那霉素抗性植株,抗性植株经PCR及PCR-Southern杂交检测,有13株均呈阳性,证明Anti-PLDγ基因成功整合到美洲黑杨G2基因组中.耐盐性实验表明,4株转基因植株抗NaCl能力比对照有不同程度提高.更多还原

【Abstract】 Antisense phospholipase Dγ(Anti-PLDγ) gene was introduced into Populus deltoides (G2) by Agrobacterium tumefaciens so as to enhance salt tolerance.Firstly,the optimal media of Populus deltoides (G2) for bud differentiation and rooting were established.The sensitivity of explants to kanamycin was tested in order to decide a reasonable selective pressure for transformation.The tranditional transformation method mediated by Agrobacterium tumefaciens was improved.21 kanamycin-resistant rooted plants were obtained through successive selection in shoot and root induction stage under high level kanamycin pressure.PCR and PCR-Southern analysis showed that 13 of 21 kanamycin-resistant rooted plants were positive.This meant that Anti-PLDγ gene was successfully integrated into the genome of these 13 plants.Salt-tolerance test demonstrated that 4/13 plants were enhanced to some extent.更多还原